The present invention relates to a novel elution complex particularly adapted to affinity chromatography and affinity precipitation.
Affinity chromatography is a technique at present very widely used for isolating enzymes and purifying them so as to obtain the desired enzyme with a high degree of purity. One of the delicate phases of this technique is the elution: the desired enzyme or sometimes an assembly of enzymes must be exclusively entrained. In fact, it has been possible to retain several types of enzymes on the chromatography column. In order to be sure that only the desired enzyme is eluted, a solution is used containing a substrate of this enzyme.
Thus, for example, the hexokinases, which have as substrate nicotinamide-adenine-dinucleotide phosphate (NADP) are eluted by means of a solution containing this compound; similarly, luciferase which has adenosine-triphosphate (ATP) as specific substrate, is eluted by means of a solution containing this acid.
Affinity precipitation is another technique which can be used for isolating enzymes and purifying them. It is closely related to affinity chromatography and immuno precipitation. This technique comprises first precipitating the enzyme to be purified with a dimerized bifunctional ligand and then specifically dissociating said enzyme with a solution containing its specific substrate.
But the substrates, particularly those mentioned above, are sometimes expensive because they are difficult to obtain. In addition, once the enzyme has been removed from the elution solution, the remaining solution cannot be reused and this recycling impossibility further increases the cost price of the enzyme.
Therefore, one of the aims of the present invention is to provide an elution complex more especially for affinity chromatography and affinity precipitation which performs as well as the specific substrate for the enzyme used to elute the enzyme retained on the chromatography column.
An object of the invention is such an elution complex, the production of which is easy and inexpensive, thus allowing enzyme production at as low a cost as possible.